Human peripheral blood-derived mononuclear cells (hPBMCs) isolated and cultured for 7–10 days, demonstrated a spindle-shaped, endothelial cell-like morphology. These differentiating cells endocytose acLDL and bind UEA-1, consistent with endothelial lineage cells.
hPBMCs were isolated by density gradient centrifugation with Histopaque-1077. Cells were plated on human fibronectin-coated culture dishes and maintained in endothelial cell basal medium-2 (EBM-2). The medium was supplemented with EGM-2 MV SingleQuots containing FBS (5%), human VEGF-1, human fibroblast growth factor-2 (FGF-2), human epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), and ascorbic acid. After 4 days in culture, nonadherent cells were removed by washing with PBS, new medium was applied, and the cells were maintained through days 7–10.